Recombinant Adenoviral Gene Transfer Does Not Affect Cardiac Allograft Vasculopathy

Background: Adenovirus serotype 5 has remained the pre-eminent vector in pre-clinical gene therapy applications in cardiac transplantation. Concerns over the potential effects of adenoviral vectors on the later development of cardiac allograft vasculopathy (CAV) are addressed in this study. Methods: Hearts (n = 22) harvested from Brown Norway rats were perfused ex vivo with either University of Wisconsin (UW) solution with no virus, Ad-CMV-LacZ or Ad-CMV-Null. Donor hearts were transplanted heterotopically into the abdomen of Lewis rats. All recipients received cyclosporine for the duration of the experiment. Transplanted hearts were recovered for analysis at 120 days. Sections of the heart were stained with elastic-van Gieson stain for morphometric analysis of the vessels to ascertain the degree of vascular luminal occlusion. Hematoxylin-eosin staining facilitated diagnosis of chronic rejection. Results: Seventy-seven percent of transplanted hearts showed signs of chronic rejection with no difference in the proportion of animals between groups (p = 0.797). No difference was noted in the degree of vascular luminal occlusion between the Ad-Null (0.57 \ub1 0.22), Ad-LacZ (0.62 \ub1 0.19) and UW (0.47 \ub1 0.29) groups (p = 0.653). Conclusions: Vascularized cardiac allografts transplanted from Brown Norway to Lewis rats demonstrated cardiac allograft vasculopathy CAV at 120 days. Adenoviral perfusion of the donor heart ex vivo did not affect the development of CAV. \ua9 2007 International Society for Heart and Lung Transplantation

Energy dysregulation and neuro-axonal dysfunction in MS measured in-vivo with diffusion-weighed spectroscopy

Neuro Imaging Researc

Energy dysregulation and neuro-axonal dysfunction in MS measured in-vivo with diffusion-weighed spectroscopy

Neuro Imaging Researc

Dysregulation of energy metabolism in multiple sclerosis measured in vivo with diffusion-weighted spectroscopy

Neuro Imaging Researc

Absolute Quantification of Aldehyde Oxidase Protein in Human Liver Using Liquid Chromatography–Tandem Mass Spectrometry

The function of the enzyme human aldehyde oxidase (AOX1) is uncertain, however, recent studies have implicated significant biochemical involvement in humans. AOX1 has also rapidly become an important drug metabolizing enzyme. Until now, quantitation of AOX1 in complex matrices such as tissue has not been achieved. Herein, we developed and employed a trypsin digest and subsequent liquid chromatography tandem mass spectrometry analysis to determine absolute amounts of AOX1 in human liver. E. coli expressed human purified AOX1 was used to validate the linearity, sensitivity, and selectivity of the method. Overall, the method is highly efficient and sensitive for determination of AOX1 in cytosolic liver fractions. Using this method, we observed substantial batch-to-batch variation in AOX1 content (21-40 pmol AOX1/mg total protein) between various pooled human liver cytosol preparations. We also observed inter batch variation in V(max) (3.3-4.9 nmol min(−1) mg(−1)) and a modest correlation between enzyme concentration and activity. In addition, we measured a large difference in k(cat)/K(m), between purified (k(cat)/K(m) of 1.4) and human liver cytosol (k(cat)/K(m) of 15-20) indicating cytosol to be 11-14 times more efficient in the turnover of DACA than the E. coli expressed purified enzyme. Finally, we discussed the future impact of this method for the development of drug metabolism models and understanding the biochemical role of this enzyme